With an approach to study the anti-tumor effects and mechanism of selenium compound, we investigated the anti-tumor activity and mechanism of $Na_5SeV_5O_{18}{\cdot}3H_2O$ (NaSeVO) in K562 cells. The results showed that $0.625{\sim}20\;mg/L$ NaSeVO could significantly inhibit the proliferation of K562 cells in vitro in a time- and concentration-dependent manner as determined by microculture tetrazolium (MTT) assay, the IC50 values were 14.41 (4.45-46.60) and 3.45 (2.29-5.22) mg/L after 48 hand 72 h treatment with NaSeVO respectively. In vivo experiments demonstrated that i.p. administration of 5, 10 mg/kg NaSeVO exhibited an significant inhibitory effect on the growth of transplantation tumor sarcoma 180 (S180) and hepatoma 22 (H22) in mice, with inhibition rate 26.8% and 58.4% on S180 and 31.3% and 47.4% on H22, respectively. Cell cycle studies indicated that the proportion of G0/G1 phase was increased at 2.5 mg/L while decreased at 10 mg/L after treatment for 24, 48 h. Whereas S phase was decreased at 2.5-5 mg/L and markedly increased at 10 mg/L after treatment for 48 h. After treatment for 24 h, 10 mg/L NaSeVO also markedly increased S and G2/M phases. Take together, the result clearly showed that NaSeVO markedly increased S and G2/M phases at 10 mg/L. The study of immunocytochemistry showed that the expression bcl-2 is significantly inhibited by 10 mg/L NaSeVO, and bax increased. Morphology observation also revealed typical apoptotic features. NaSeVO also significantly caused the accumulation of $Ca^{2+}$ and $Mg^{2+}$, reactive oxygen species (ROS) and the reduction of pH value and mitochondrial membrane potential in K562 cells as compared with control by confocal laser scanning microscope. These results suggest that NaSeVO has anti-tumor effects and its mechanism is attributed partially to apoptosis induced by the elevation of intracellular $Ca^{2+}$, $Mg^{2+}$ and ROS concentration, and a reduction of pH value and mitochondria membrane potential (MMP).
Park, Gil-Soon;Chang, In-Ae;Kim, Youn-Chul;Lee, Moo-Hyung;Shin, Hye-Young;Choi, Du-Young;Yun, Yong-Gab;Park, Hyun
Journal of Physiology & Pathology in Korean Medicine
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v.21
no.3
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pp.700-704
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2007
In the recent, increased concern has been focused on the pharmacology and clinical utility of herbal extracts and derivatives as a drug or adjunct to chemotherapy and immunotherapy. Here we investigated the role of the extract of Anethi Fructus in the expression of inflammatory mediators, surface molecule, and related receptors in vitro. In murine macrophage RAW 264.7 cells and peritoneal macrophages of C57BL/6N mice, water extract of Anethi Fructus increased the production of secretary tumor necrosis factor (TNF)-a and Nitric oxide (NO), and the expression level of CD14, LPS co-receptor and CD86, co-stimulatory molecule compared to negative natural extract ex vivo. The water extract of Anethi Fructus increased the production of interferon (IFN)-g from splenocytes. Also, water extract of Anethi Fructus increased ConA-induced cell proliferation. These results suggest that water extract of Anethi Fructus may enhance the immune response through immune modulation of macrophage and lymphocytes.
Objectives: This study was designed to investigate the anti-tumor metastasis by anti-inflammatory and innate immunomodulating effects of extracts of Jipae-san on cancer cells. Methods: Antimetastatic experiments were conducted in vivo mouse model by using 4T1 mouse mammary carcinoma cells. Cell viability of Jipae-san was tested with 4T1 mouse mammary carcinoma cells, colon 26-M3.1 carcinoma cells and macrophage. In addition expression of $TNF-{\alpha}$ and NO induced by LPS was measured after treating with Jipae-san. To observe innate immunomodulating effects of Jipae-san on macrophage, we measured $TNF-{\alpha}$, IL-12, IL-6 and MCP-1, respectively. Cell cytotoxicity was tested with the macrophage stimulated with Jipae-san and we evaluated the activation of $TNF-{\alpha}$ and NO. And the effect of Jipae-san on metastasis was measured without NK-cell using GM1 serum. Results: Intravenous inoculation of Jipae-san significantly inhibited metastasis of 4T1 mouse mammary carcinoma cells. In an in vitro cytotoxicity analysis, cell growth are closer to 100% less than $1,000{\mu}g/ml$ concentration. The expression of $TNF-{\alpha}$ and NO induced by LPS after treating Jipae-san was down regulated in dose-dependent manner. Level of cytokines such as $TNF-{\alpha}$, IL-12, IL-6 and MCP-1 of Jipae-san group were up regulated in compared to the control group. The macrophage stimulated with Jipae-san significantly inhibits the cancer cell at ratio of 10:1, 20:1. The activation of NO was significantly up regualted in a group of 5:1, 10:1, 20:1. The depletion of NK-cells by anti-asialo GM1 serum partly abolished the inhibitory effect of Jipae-san on tumor metastasis. Conclusions: Jipae-san appears to have considerable activity on the anti-metastasis by inflammation control and activation of innate immune system.
Hwang, Buyng Su;Lee, Seung Young;Kang, Chang Hee;Han, Woog;Oh, Young Taek;Yu, Sang Mi;Kim, Min Jin;Kim, Chul Hwan;Eom, Jung Hye;Jeong, Sang Chul;Lee, Wook Jae;Ahn, Young Hee;Jeong, Yong Tae
Microbiology and Biotechnology Letters
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v.47
no.2
/
pp.195-200
/
2019
The purpose of this study was to investigate the melanogenesis inhibiting activity of the ethanol extract from Polygonum amphibium L. Firstly, the n-hexane (Hx), chloroform ($CHCl_3$), ethyl acetate (EA), n-butanol (BuOH), and water (Water) fractions were isolated from the P. amphibium L. ethanol extract. The efficacy of melanogenesis was found to significantly decrease via the EA and BuOH fractions when compared to the control in B16F10 cells. EA particularly showed the lowest melanin content in B16F10 cells when compared to all the other extracts. Concentration-dependent inhibition of melanin synthesis was also observed in the EA fraction at concentrations below $50{\mu}g/ml$, which did not exhibit cytotoxicity in B16F10 cells. Notably, the expression of three key proteins (tyrosinase, tyrosinase-related protein-1 (TRP-1), and TRP-2), which are involved in melanogenesis, were significantly decreased via the EA fraction. EA also inhibited body pigmentation in vivo in a zebrafish model. Overall, we demonstrated melanogenesis suppression using the EA fraction from P. amphibium L., which could be a potential candidate for an antimelanogenesis agent.
Purpose: This study examined the antioxidant and cancer cell growth inhibitory activities of an ethanol extract and different solvent fractions of Mesembryanthemum crystallinum L. (ice plant). Methods: The ice plant was freeze-dried, extracted with 99.9% ethanol, and then fractionated with hexane, ethyl acetate, butanol, and water. The total polyphenol content (TPC), total carotenoid content (TCC), 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity (RSA), and ferric reducing antioxidant power (FRAP) were measured. Assays using 2',7'-dichlorofluorescin-diacetate and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide were performed to measure the intracellular reactive oxygen species (ROS) and cell growth, respectively. Annexin V/propidium iodide staining and cell cycle analysis were performed for the detection of apoptosis and cell cycle arrest. Results: TPC, TCC, RSA, and FRAP of the ethanol extract (EE) were 3.7 mg gallic acid equivalent/g, $13.2{\mu}g/g$, 21.0% (at a concentration of 5 mg/mL), and 21.0% (at a concentration of 5 mg/mL), respectively. Among the different solvent fractions, the butanol fraction (BF) showed the highest TPC (5.4 mg gallic acid equivalent/g), TCC ($86.6{\mu}g/g$), RSA (34.9% at 5 mg/mL), and FRAP (80.8% at 5 mg/mL). Treatment of HCT116 human colon cancer cells with EE and BF at concentrations of 250 and $500{\mu}g/mL$ reduced the levels of intracellular ROS. Concomitantly, EE and BF resulted in the dose-dependent inhibition of cell growth (at the concentrations of 125, 250, and $500{\mu}g/mL$ for 24 ~ 48 h) and the induction of apoptosis (at the concentrations of 250 and $500{\mu}g/mL$ for 48 h) in HCT116 cells. An increased G2/M cell population was also found in the BF-treated cells. Conclusion: These results suggest that ice plant possesses antioxidant and growth inhibitory activities in colon cancer cells.
Proceedings of the Plant Resources Society of Korea Conference
/
2019.10a
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pp.92-92
/
2019
Excessive or chronic inflammation contributes to the pathogenesis of many inflammatory diseases such as sepsis, rheumatoid arthritis, and ulcerative colitis. Hibiscus syriacus L. has been used as a medicinal plant in many Asian countries, even though its anti-inflammatory activity has been unclear. Therefore, we investigated the anti-inflammatory effect of anthocyanin fractions from the H. syriacus L. varieties Pulsae (PS) on the lipopolysaccharide (LPS)-induced expression of proinflammatory mediators and cytokines in RAW264.7 macrophages. PS suppressed LPS-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) secretion concomitant with downregulation of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. Furthermore, PS inhibited the production of proinflammatory cytokines such as tumor necrosis factor-alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), and IL-12 in LPS-stimulated RAW264.7 macrophages. Further study showed that PS significantly decreased LPS-induced nuclear translocation of the nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) subunits, p65 and p50. Molecular docking data showed that many anthocyanins from PS fit into the hydrophobic pocket of MD2 and bound to Toll-like receptor 4 (TLR4), indicating that PS inhibits the TLR4-MD2-mediated inflammatory signaling pathway. Especially, apigenin-7-O-glucoside most powerfully bound to MD2 and TLR4 through LYS122, LYS122, and SER127 at a distance of $2.205{\AA}$, $3.098{\AA}$, and $2.844{\AA}$ and SER441 at a distance of $2.873{\AA}$ (docking score: -8.4) through hydrogen bonding, respectively. Additionally, PS inhibited LPS-induced TLR4 dimerization/expression on the cell surface, which consequently decreased MyD88 recruitment and IRAK4 phosphorylation. PS completely blocked LPS-mediated mortality in zebrafish larvae by diminishing the recruitment of neutrophil and macrophages accompanied by low levels of proinflammatory cytokines. Taken together, our results indicate that PS attenuates LPS-mediated inflammation in both in vitro and in vivo by blocking the TLR4/MD2-MyD88/IRAK4-$NF-{\kappa}B$ axis. Therefore, PS might be used as a novel modulatory candidate for effective treatment of LPS-mediated inflammatory diseases.
Hwang, Mi Sun;Lee, Joo Hee;Kim, In Joo;Kim, Tae Hee;Kim, Kang Min
Journal of Life Science
/
v.30
no.9
/
pp.791-797
/
2020
Maqui berry are known as one of the "superfoods" and can purportedly improve health. The effects of orally administered maqui berry extract on biological parameters such as serum lipids and blood cells in healthy subjects were investigated in rats in vivo for 2 weeks. Possible causes of harmful effects on liver and kidney function were also examined. After oral administration of maqui berry extract, levels of serum triglyceride (TG) were decreased to almost 30% less from the control, and high density lipoprotein cholesterol (HDL-C) was increased to 18% more; thus, serum lipids profile seemed to be improved. The administration of maqui berry extract increased red blood cell concentration by 10% from the control, while both the hemoglobin and the mean corpuscular volume (MCV) were increased by 12%. Maqui berry extract also increased the concentration of serum total proteins by 30% of the control and that suggests protein metabolism might be improved. Moreover, maqui berry decreased serum levels of glutamin oxaloacetate transaminase (GOT) and glutamin pyruvate transaminase (GPT) by 12% from the control group so maqui berry improved liver function. This study suggests that oral administration of maqui berry extract may be helpful to improve those heath parameters like serum lipids profile, red blood cell level, liver protection. It would also be very useful for the development of high value products such as functional foods and pharmaceutics in the future.
Park, Sung Il;Park, Ju Hyung;Lee, Chi Kug;Kim, Shin Chae;Choi, Bo Geum;Kwak, Jae Yong;Yim, Chang Yeol
IMMUNE NETWORK
/
v.1
no.2
/
pp.162-169
/
2001
Background: Nitric oxide (NO), a cytotoxic molecule is produced in various tissues including tumor cells during interleukin-2 (IL-2) therapy . Lymphokine-activated killer (LAK) cells are induced during IL-2 therapy, and have cytotoxic activity against tumor cells. The current study investigated the effects of NO synthesized in target cells or exposure of target cells to NO on the sensitivity of target cells to LAK cell cytotoxicity. Methods: Cytotoxicity was measured using 4 h chromium release assays. LAK cells which were induced by a 4 day incubation of BALB/c mouse splenocytes with IL-2 (6,000 IU/mL) were employed as effector cells. RD-995 skin tumor cells originated from a C3H/HeN mouse were employed as target cells. NO synthesis in target cells was induced by a 24 h incubation of RD-995 cells with $IFN{\gamma}$ (25 U/mL), TNF (50 U/mL) and IL-1 (20 U/mL). S-nitrosyl acetylpenicillamine (SNAP), an NO donor, was used to expose target cells to NO. $N^G$-monomethyl-L-arginine (MLA) and carboxy-PTIO were added during cytotoxicity assays to inhibit NO synthesis, and to scavenge NO produced by target cells, respectively. Results: Sensitivity of NO-producing RD-995 cells to LAK cell cytotoxicity was decreased by addition of MLA and carboxy-PTIO during cytotoxicity assays. However, the two reagents had no effect on the sensitivity of non-NO-producing RD-995 cells. Pretreatment of RD-995 target cells with SNAP increased the sensitivity in comparison with untreated cells. Conclusions: Sensitivity of target cells to LAK cell cytotoxicity is increased by target cell NO synthesis or exposure to NO. Further studies are needed to evaluate whether these in vitro results have relevance to in vivo phenomena.
Journal of the Korean Society of Food Science and Nutrition
/
v.42
no.11
/
pp.1744-1752
/
2013
We investigated the protective effect of UVB inducing photodamage from mulberry extract (ME) and Lithospermum erythrorhizon extract (LE). The contents of total anthocyanin and shikonin as a color compound of ME and LE were 4.92 mg/g and 9.58 mg/g, respectively. The electron donating ability and superoxide radical scavenging activity of ME were 84.32% and 76.34%, respectively. The oxygen radical absorbance capacity of the ME ($545.37{\mu}moles$ TE/g) was higher than LE ($427.18{\mu}moles$ TE/g). MMP-1 production in the HS68 cells were exposed to UVB suppressed by treatment with $200{\mu}g/mL$ of ME (68.6%) and LE (32.7%). ME and LE were applied to a skin aging mouse model, which was induced by the irradiation of UVB to the backs of hairless mice. The value of skin erythema index, wrinkle depth and thickness, epidermis thickness, and collagenous fiber damage in the experiment groups (MEL: ME 3%, MEM: ME 5%, MEH: ME 7%, LEL: LE 3%, LEM: LE 5%, LEH: LE 7%) were remarkably reduced than in the control group (only UVB exposure group), while water capacity increased. The level of total wrinkles depth in the skin was decreased to be 30% of the control group by MEH and LEM. These results suggest that ME and LE are useful cosmetic materials for skin protection against UVB-inducing.
Capsaicin is known to be an analgesic agent, affecting the synthesis, storage, , transport and release of substance p, the principal neurotransmitter of pain from periphery to the central nervous system(CNS). DA-5018, a newly synthesized capsaicin derivative has shown potent analgesic effect comparable to that of morphine in various rat models of experimentally inducted acute pairs. In this study the mechanism of analgesic actlvity of DA-5018 was examined. First, the electrically-evoked contraction of guinea pig trachea was inhibited by DA-5018 and these inhibition was recovered by incubation with capsafepine(3$\muM$), capsaicin receptor antagonist and this result suggested that DA-5018 has affinity on capsaicin receptor. The correlation between the norciceptive threshold and the release of substance P was evaluated. In vivo perfusion of slices of the rat spinal cord with DA-5018(10, 100$\muM$) produced a significant increase of the release of substance P and this increase was less than that of capsaicin(10$\muM$). The norciceptive threshold of rat treated with DA-5018(1 mg/kg, p.o) in tall pinch test increased from 2.9$\pm$0.3 to 23.5 $\pm$6.61. Tail pinch latency increased to a maximun at 15 min after DA-5018 treatment and then declined to control values by 120 min. The capsaicin-evoked release ot substance P from the spinal cord slices of rat treated with DA-5018 reduced from 2.38$\pm$ 0.79 to 0.69$\pm$ 0.26 pg/mg wet weight. This reduction reached to a minium at 15 min after DA-5018 treatment and then recovered to control value by 120 min. These results mean that analgesic activity of DA-5018 is due to release of substance P The effect of DA-5018 cream on electrically-evoked neurogenic inflammation of rat saphenous nerve was compared with capsaicin (zostrix-HP). DA-5018 showed 34% inhibition of the neurogenic extravasation while capsaicin showed significant 67% inhibition. This result indicates that the potency of DA-5018 in the release of substance P is less than that of capsaicin. These results suggest that the release of substance P is partially involved in the mechanism of analgesic action of DA-50l8.
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